膀胱上皮细胞的体外原代培养方法

摘要

BACKGROUND: There are three methods to obtain human bladder epithelial cells: enzymatic digestion, scraping and tissue plant. Each of the three methods has its advantage and disadvantage. There are too many ingredients in the culture medium to deal with, besides, the efficiency is low.OBJECTIVE: To approach the best method for primary culture of human bladder epithelial cells.METHODS: Human bladder epithelial cells from normal bladder mucosa were cultured with three different primary cell culture methods: enzymatic digestion, scraping and tissue plant.RESULTS AND CONCLUSION: The successful rates of bladder epithelial cells cultured with enzymatic digestion, scraping and tissue plant were 13.3%, 26.7% and 86.7% respectively, and the differences in successful rates were significant (P < 0.001). Fluorescent staining with AE1/AE3 antibody confirmed that the cultured bladder epithelial cells were positive for AE1/AE3 epithelial cytokeratins. Tissue plant is a simple and feasible primary culture method for bladder epithelial cells culture, and a great amount of pure bladder epithelial cells can be obtained in a short time using this method.%背景:国内外获得原代人正常膀胱上皮细胞的方法包括酶消化培养法、刮削培养法、组织块培养法,这些方法各有优缺点,培养基中成分多不易把握,效率不高.目的:分析人膀胱上皮细胞原代培养的最佳方法.方法:在同一实验条件下,采用酶消化培养法、刮削培养法、组织块培养法3种不同的原代细胞培养方法培养人正常膀胱黏膜膀胱上皮细胞.结果与结论:酶消化培养法、刮削培养法、组织块培养法各组膀胱上皮细胞培养成功率分别为13.3%,26.7%,86.7%,3组间比较差异有非常显著性意义(P < 0.001).3组培养的人膀胱上皮细胞角蛋白AE1/AE3荧光染色均呈阳性.说明组织块培养法是一种简单,短期内可扩增出较多纯净的膀胱上皮细胞的原代培养方法.